Philippine Science Letters
vol. 4 | no. 2 | 2011
published online December 16, 2011


Microsatellite and RAPD Analysis of Six Local Wine Strains of Saccharomyces cerevisiae

by Zahara Joy A. Guiamal and Cynthia T. Hedreyda*

National Institute of Molecular Biology and Biotechnology, College of Science, University of the Philippines, Diliman, Quezon City, Philippines 1101



The development of fast and reliable molecular typing techniques that can identify genetic markers specific for industrially important strains of Saccharomyces cerevisiae are valuable for wine fermentation and biofuel production and can ensure the use of correct strains and will pave the way for producing consistent quality of product. Preliminary experiments verified that five (WS-1, WS-3, WS-4, WS-5, WS-6) of six local wine strains of Saccharomyces cerevisiae used in this study exhibited better growth than the control non-wine strain in the presence of 5% ethanol, while five (WS-1, WS-2, WS-3, WS-5, WS-6) of six strains grew better than the control in 10% ethanol. Growth of four wine strains (WS-1, WS-2, WS-5, and WS-6) was comparable to the control while two strains (WS-3 and WS- 4) exhibited significantly higher growth in 15% ethanol. All the yeast strains generated an identical DNA profile from PCR using the microsatellite primer (GAC)5 except strain WS-4 and the control that generated unique profiles. Results of randomly amplified polymorphic DNA (RAPD) PCR on the yeast strains revealed that 8 of the 11 primers were able to distinguish all the wine strains from the control strain. RAPD PCR using the M13 primer and three 10-mer RAPD primers (OPA-11, OPY-02, and OPY-05) resulted in profiles unique to strain WS-4 and the control non-wine strain. RAPD primer 1283 generated unique profiles for WS-3 in addition to WS-4 and the control. Additional unique RAPD profiles were also observed for wine strain WS-3 with PCR primers OPY-3 and OPY-4, while WS-2 produced a unique profile with primer OPB-11. The significant difference in the ethanol tolerance of control versus wine strains, strain WS-4 versus other strains, as well as some unexpected ethanol tolerance results for strains WS-2 and WS-3, could be attributed to significant genetic variability of the strains that was also reflected as variations in DNA profiles generated through the microsatellite and several RAPD PCR. Results suggest that the molecular typing tools used in this study may be able to generate DNA profiles that could distinguish S. cerevisiae strains exhibiting different ethanol tolerance.

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Submitted: July 1, 2011
Revised: October 5, 2011
Accepted: October 5, 2011
Published: December 16, 2011
Editor-in-charge: Michael D. Purugganan
Michael D. Purugganan